Download Spectrophotometric Microdetermination of Anthelmentic Drug in Pure Form

Journal of Applied Sciences Research, 4(10): 1242-1248, 2008
© 2008, INSInet Publication
Spectrophotometric Microdetermination of Anthelmentic Drug in Pure Form
and Pharmaceutical Formulation by Ion-pair Complexation
1
Manal S. Kamel, 2 B.N.Barsoum and 2 Rania Sayed
1
Department of Applied Organic Chemistry, Microanalytical Lab.,
National Research Centre, Cairo, Egypt.
2
Department of Chemistry, Faculty of Science, Cairo University, Cairo, Egypt.
Abstract: Simple, sensitive and rapid spectrophotometric procedures are described for the quantitation
determination of anthelmentic drug (albendazole). The procedures are based on the ion-pair complexation
reactions. In the procedure the examined drug (albendazole) was reacted with thymol blue and alizarin
derivatives., alizarin (I), alizarin red S (II), alizarin yellow (III), quin alizarin (IV), alizarin fluorine blue
(V) and thymol blue (VI) in distilled water producing coloured ion-pair complexes which can be
measured at the optimum wavelength for each complex. The optimization of the reaction conditions is
investigated. Beer's law is obeyed in the concentration ranges 6.6325 - 92.855 µg ml-1. The molar
absorptivity is also calculated. The correlation coefficient was $ 0.98 (n = 5), with a relative standard
deviation (R.S.D) of # 2.67 for five determinations. The methods are successfully applied to the
determination of albendazole in their pharmaceutical formulation. The results obtained from
pharmaceutical preparations compared well with those obtained by the official method [1] and
demonstrated good accuracy and precision.
Key words: Spectrophotometric, albendazole, alizarin derivatives, thymol blue and pharmaceutical
formulation.
INTRODUCTION
Albendazole is chemically methyl-5-(propylthio)-2benzimidazolecarbamate, C 12H 15N 3O 2S [1]. It is a widespectrum anthelmentic drug used for human and animal
infections. W hen administered orally, it is quickly
biotransformed into its active intermediate metabolite
albendazole-sulphoxide (ABZSO), which is then
oxidized to the inactive form of albendazole-sulphone
(ABZSO 2). Because of their affinity for the parasite âtubulin, both alb end azo le and AB ZSO show
anthelmentic activity [2]. It is also widely used as
inhibator for protein synthesis and causes degenerative
changes in the intestine and the enveloping membrane
and for treatment of cysticercosis has received
criticisms [3,7], their use in the cysticidal treatment of
neurocysticercosis has proved efficacious [8,9]. Although
neurocysticercosis is widespread in underdeveloped
countries with an important socioeconomical impact,
there is a lack of information about its natural history,
treatment and prognosis [3,10] and widely used for
treatment and control of helminthes in cattle [11,12].
Several methods have been reported for the
determination of albendazole including HPLC [13,14], IonPair Liquid Chromatography[15], HPLC-electrospray
m a s s s p e ctro m e try [ 16] , sp e c tr o p h o to m e tr y [1 7 , 1 9 ] ,
titrimetry[18,19] and FIA [20].
In order to continue our work for using alizarin
derivatives and thymol blue for drug analysis, a
simple, accurate, economic, sensitive, more essential
and less-time consuming spectrophotometric method for
the determination of the anthelmentic drug under
investigation in pure and in their dosage form are
performed.
This paper describes the application of ion-pair
com plexation reactions for sp ectro photom etric
determination of albendazole in pure form and
pharmaceutical preparation.
Corresponding Author: Manal S. Kamel, Department of Applied Organic Chemistry, Microanalytical Lab., National
Research Centre, Cairo, Egypt.
E-mail: [email protected]
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J. Appl. Sci. Res., 4(10): 1242-1248, 2008
flask.
M ATERIALS AND M ETHODS
Apparatus: A SHIM ADZU UV 160-A is a double
beam UV-Visible recording spectrophotometer with a
10 mm quartz cell was used for all spectrophotometric
measurements, a HANA microprocessor pH meter 8417
was used for checking the pH of buffer solutions.
M aterials: Albendazole stock solution (10 -3M), was
prepared by dissolving 0.02653 g of albendazole in
small amount of anhydrous formic acid in 100 ml
measuring flask then completed with distilled water to
the mark.
Amoun pharmaceutical Company, El-Obour City,
Cairo, Egypt supplied albendazole in pure form and
their pharmaceutical formulation, [vermizole syrup (it
was labeled to contain 30 ml where each 5 ml is
equivalent to 200 mg albendazole per tablet) Batch
NO.: 3696].
Reagents: All the reagents and solvents used were of
analytical grades. All solutions were freshly prepared.
C
Alizarin, 1,2-dihydroxyanthraquinone (I), a stock
solution (10 -4 M) was prepared by dissolving
0.0012 g in slightly alkaline media (2M NaOH)
then completed by distilled water to 50 ml in a
measuring flask.
C
Alizarin red S, 9,10-dihydro-3,4-dihydroxy-9,10dioxo-2-anthracenesulfonic acid sodium salt (II), a
stock solution (10 -3 M) was prepared by dissolving
0.0179 g in 50 ml distilled water in a measuring
flask.
C
Alizarin yellow G,5-(4-nitrophenylazo)salicylic acid
sodium salt (III) , a stock solution (10 -3 M) was
prepared by dissolving 0.01546 g in 50 ml distilled
water in a measuring flask.
C
Q uina liz arin,
1 ,2,5,8-tetrahyd ro xy-9 ,1 0 anthraquinone (IV), a stock solution (10 -4 M) was
prepared by dissolving 0.00136 g in slightly
alkaline media (2M NaOH) then completed by
distilled water to 50 ml in a measuring flask.
C
Alizarin fluorine blue, 3,4-dihydroxyanthraquinon2-yl-methylimino-diacetic acid (V), a stock solution
(10 -3 M) was prepared by dissolving 0.019267 g in
50 ml distilled water in a measuring flask.
C
Thymol blue (VI), thymolsulfonphthalein, a stock
solution (10 -4 M) was prepared by dissolving
0.0023 g in 50 ml distilled water in a measuring
C
2M NaOH solution.
C
A buffer solution of pH 3, was prepared by mixing
6 ml of 2M sodium hydroxide solution with 10 ml
of 2 M citric acid solution and diluting to 100 ml
with distilled water.
A series of buffer solution of different pH were
adjusted by NaOH solutions.
BDH supplied alizarin deravatives which made in
England and finchemie K.-H.Kallies KG supplied
thymol blue which made in Germany.
General Procedure: 1.0 ml of albendazole solution
(10 -3M) was added to 1.0 ml of reagents (I,III,IV,VI)
and transferred into 10 ml measuring flask and
completed to the mark with distilled water. For reagents
(II ,V) 1.0 ml of albendazole solution
(10 -3M) was mixed with 1.0 ml of reagents then add
5.0 ml of the buffer solution (citrate) of the optimum
pH values as recorded in (Table 1) and completed to
10 ml with distilled water in measuring flask then for
reagent (II) diluted by 1:7 with distilled water, the
absorbance was measured at the optimum wavelength
(Table 1) against a water blank.
Application to Various Dosage Forms: For vermizole
syrup (albendazole) filterate it and take 0.75 ml (0.75
ml is equivalent to 30 mg) of the clear solution and
then completed to 100 ml with distilled water in
measuring flask . the general procedure is applying to
the drug content of this solution.
The results are obtained similar to the results in
the official method [1].
RESULTS AND DISCUSSION
Optimization: Careful investigations were carried out
to establish the most favorable conditions to achieve
maximum colour intensity in the quantitative
determination of the examined anthelmentic drug
(albendazole). The absorption spectra of albendazole
and their complexes with alizarin derivatives (I-V) and
thymol blue (VI) under the optimum conditions are
shown in figures (1 - 6) and recorded in (Table 1), the
absorption band of albendazole complexes are located
at 294, 320, 352, 282, 502 and 291 nm with reagents
(I-VI), respectively.
However, in all instances the absorbance was
measured at this ë max against a water as a blank under
identical conditions. The influence of each of the
following variables on the reaction was tested.
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J. Appl. Sci. Res., 4(10): 1242-1248, 2008
Table 1: Quantitative parameters for the complexation of albendazole with alizarin derivatives (I-V) and thymol blue (VI).
parameter
I
II
IIII
IV
V
VI
ëm ax (nm)
294
320
352
282
502
291
--------------------------------------------------------------------------------------------------------------------------------------------------------------------------------pH
7
9
9
9
9
9
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------reagent(ml)
2.5
2
2
1.5
1.5
2
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------time (min.)
20-50
40-60
30-60
0-40
10-60
0-50
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Beer's law limit (µg/ml)
26.53-53.06
26.53-79.59
13.265-39.795
13.265-66.325
26.53-92.855
6.6325-79.59
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------molar absorptivity
0.158
0.006
-0.254
0.137
0.013
0.159
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------sandell sensitivity
1.004×103
0.367×103
0.508×103
1.99×103
2.203×103
1.07×103
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Standard divition
0.264
0.523
0.133
0.12
0.72
0.248
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Specific absorptivity
2.3×10-5
1×10-6
-3.8×10-5
2×10-5
1×10-6
2.4×10-5
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Regression equation
Y = ab + c
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Slope (a)
0.023
0.001
-0.038
0.020
0.001
0.024
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Intercept (c)
0.364
0.011
2.356
0.148
0.038
0.118
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Correlation
0.997
0.985
0.980
0.991
0.991
0.996
coefficient (r)
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------R.S.D. (%)
0.95
0.488
0.48
2.67
8.66
1.798
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------Range of error
0.522
1.4
0.368
0.3
1.9
0.47
Scheme 1: proposed reaction pathway between albendazole and R, R is a reagent (I-VI).
The reaction mechanism of albendazole with
alizarin derivatives (I-V) and thymol blue (VI) was
proposed in scheme 1.
Effect of pH: Series of citrate buffer of pH values
(3-12) were examined to achieve maximum colour
intensity. The optimum pH were 7,9,9,9,9 and 9 for
reagents (I-VI), respectively as shown in (Table 1).
Effect of the Reagent Concentration: The effect
ofreagent was investigated by taking various amount of
reagent added to an aliquot of solution containing 1 ml
of drug under investigation (albendazole) and follow of
the procedure of each reagent, the volume of reagent
was increased from 0.5 to 3.5 ml. The maximum
absorption was observed with the addition of 2.5 ml for
reagent (I), 2 ml for reagents (II,III,VI) and 1.5 ml for
reagent (IV,V) as shown in (Table 1).
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J. Appl. Sci. Res., 4(10): 1242-1248, 2008
Effect of Time and Temperature: The optimum
reaction time was determined by following the colour
intensity. For reagent (I) the absorbance was
Fig. 1:
A b s o rp tio n
s p ectra
o f alb e n d a z o le
complexed with alizarin against water
blank.
Fig. 2:
A b so r p ti o n
s p e c tr a
o f a lb e nd a z o le
complexed with alizarin red S in buffer
solution of pH=7 against water blank.
Fig. 3:
A b so rp tio n
s p e c tr a
o f a lb e n d a z o le
complexed with alizarin yellow against
water blank.
increased with time consuming and stable in the period
20-50 min., for reagent (II) the absorbance is stable in
the period 40-60min., for reagent (III) the absorbance
was decreased with time consuming and stable in the
period 30-60 min., for reagent (IV) the absorbance
Fig. 4:
A b so rp tio n
s p e c tr a
o f a lb e n d a z o le
complexed with quinalizarin against water
blank.
Fig. 5:
A b s o rp tio n
s p e c tra
of a lb e nd a z o l e
complexed with alizarin flourine blue in
buffer solution of pH=7 against water
blank.
Fig. 6:
A b so rp tio n
s p e c tr a
o f a lb e n d a z o le
complexed with thymol blue against water
blank.
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J. Appl. Sci. Res., 4(10): 1242-1248, 2008
Fig. 7:
Linearity of absorbance to concentration of
albendazole with alizarin.
Fig. 8:
Linearity of absorbance to concentration of
albendazole with alizarin red S.
Fig. 11: Linearity of absorbance to concentration of
albendazole with alizarin fluorine blue.
Fig. 12: Linearity of absorbance to concentration of
albendazole with thymol blue.
stable in the period 0-40 min., for reagent (V) the
absorbance was increased with time consuming and
stable in the period 40-50 min., for reagent (VI) the
absorbance was decreased with time consuming and
stable in the period 10-60 min.,the maximum colour
development have the optimum time as shown in
(Table 1). It is clear that all conditions studied were
optimized at room temperature (25 ± 4 oC ).
Linearity of absorbance to concentration of
albendazole with alizarin yellow.
Sequence of Addition: The optimum sequence was
defined by following the colour intensity and maximum
absorbance on changing the sequences of addition of
drug, reagent and buffer was shown that all sequences
give the same absorbance.
Fig. 10: Linearity of absorbance to concentration of
albendazole with quinalizarin
Interference: For the determination of albendazole with
reagent (I) there are an interference was observed from
the presence of Starch, Sodium bromide, Citric acid
monohydrate, Ferric chloride, Cobalt (II) chloride,
Calcium (II) chloride, and no interference observed
from the presence of Nickel (II) chloride, Sodium
sulphate anhydrous, Lactose, Ammonium chloride,
Urea, D-Glucose, Sucrose. For reagent (II) there are an
interference was observed from the presence of Citric
acid monohydrate, Nickel (II) chloride Ammonium
chloride, Starch, Cobalt (II) chloride, Sodium sulphate
anhydrous, Lactose, D-Glucose, Ferric chloride,
Calcium (II) chloride, Urea, Sucrose, and no
Fig. 9:
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J. Appl. Sci. Res., 4(10): 1242-1248, 2008
Table 2: Analysis of albendazole with alizarin derivatives (I-V) and thymol blue (VI).
Reagent
Found (µg/ml)
Taken (µg/ml)
R (%)
±S.D.
I
28.2
28.25
99.8
0.16
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------II
39.16
39
100.4
0.51
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------III
26.59
26.67
99.7
0.198
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------IV
29.347
29.33
100.05
0.05
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------V
27.204
27.2
100.015
0.12
---------------------------------------------------------------------------------------------------------------------------------------------------------------------------------VI
30.96
31
99.87
0.14
interference observed from the presence of Sodium
bromide. For reagent (III) there are an interference was
observed from the presence of Sodium bromide, Citric
acid monohydrate, Nickel (II) chloride, Starch, Cobalt
(II) chloride, Sodium sulphate anhydrous, Lactose,
Ferric chloride, Calcium (II) chloride, Urea, Sucrose,
and no interference observed from the presence
Ammonium chloride, D-Glucose.
For reagent (IV) there are an interference was
observed from the presence of Sodium bromide, Citric
acid monohydrate, Nickel (II) chloride, Starch, Cobalt
(II) chloride, Sodium sulphate anhydrous, Lactose,
Ferric chloride, Calcium (II) chloride, Urea, Sucrose,
Ammonium chloride, and D-Glucose i.e. there are
interference observed from the presence all the previous
substances. For reagent (V) there are an interference
was observed from the presence of Sodium bromide,
Citric acid monohydrate, Nickel (II) chloride, Starch,
Cobalt (II) chloride, Sodium sulphate anhydrous, Ferric
chloride, Sucrose, Ammonium chloride, and D-Glucose
and there are no interference observed from the
presence of Calcium (II) chloride, Urea, Lactose.
For reagent (VI) there are an interference was
observed from the presence of Citric acid monohydrate,
Starch, Ferric chloride, Calcium (II) chloride, Urea and
there are no interference observed from the presence of
Sodium bromide, Nickel (II) chloride, Cobalt (II)
chloride, Sodium sulphate anhydrous, Lactose Sucrose,
Ammonium chloride, and D-Glucose. The results
indicate that up to 100-fold excess of them which may
present in its pharmaceutical preparations (in case of
non interference absorbance changes by ± 3.0% which
is non - interference).
Analytical Applications: The proposed method was
successfully applied to the dosage form syrup
albendazole (vermizole). The results are recorded in
(Table 2) compared statistically with the official
method [1] reveal that the recoveries are in the range
(100.4 - 99.66) reflecting a high accuracy, in addition
to the high precision indicated very law values of
relative standard deviations. Therefore, it can be
concluded that the results of the present method are in
high agreement with those obtained by the official
method [1].
Conclusion: Alizarin derivatives and thymol blue are
a suitable reagents for the determination of anthelmentic
drugs such as albendazole in pure form or in its dosage
forms. The suggested method is simple, time saving,
sensitive and reproducible. Therefore the proposed
method can be used advantageously as a routine
method for the determination of albendazole in quality
control and industry.
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